Dr. Sachin Kumar Singh Chauhan, Ph.D.

• Certainly! Here are the key points for the resume:
  
  1. **Tissue Biopsy Processing**: Isolated tumor-infiltrated lymphocytes (T cells, NK cells, and ILCs) and prepared snap-frozen and FFPE tissue blocks.
  2. **Spectral Flow Cytometry**: Analyzed tumor-infiltrating lymphocytes using 31-color FCM panels on Cytek Aurora.
  3. **Single-Cell Analysis**: Multiplexed patient samples and synthesized cDNA libraries with BD Rhapsody, using AbSeq/TotalSeq antibodies.
  4. **Sequencing Preparation**: Prepared single-cell whole transcriptome mRNA and AbSeq libraries for scRNA sequencing.
  5. **Technical Proficiency**: Utilized Unix OS, HPC, R software (Seurat), Cellinics, BD SeqGeq software, and data modeling; mentored students and collaborated on multidisciplinary research.

• **Therapy Development**: Developed GMP-compliant NK cell-based therapy for resistant HNSCC cells using CliniMACS Prodigy.
• **Genetic Modification**: Conducted genetic modifications of NK cells to enhance efficacy.
• **Immune Checkpoint Studies**: Investigated the role of immune checkpoints in NK cell functionality and efficacy.
• **Cytokine Influence Studies**: Used multi-color flow cytometry and cell sorting to explore cytokine influences on NK cells.
• Proficient in diverse cellular and molecular techniques including tissue culture, flow cytometry, cell sorting, immunoprecipitation, cytokine assays, qRT-PCR, cell proliferation and functional assays, retroviral vector transduction, and electroporation.
• Researched gene modification methods for primary NK cells at Fraunhofer-Institut für Zelltherapie und Immunologie, Leipzig, Germany, using lentiviral and retroviral vectors encoding reporter genes.
• Completed secondment at Miltenyi Biotec, Germany, conducting live-cell cytox assays with human NK cells against solid tumors using Incucyte, and functional assays with MACS Quant flow cytometry and MACSima imaging for real-time tumor cell death measurements.
• **Regulatory Compliance and Lab Management**: Acquired proficiency in GMP quality control, accurate documentation, SOP development, lab safety, equipment setup, inventory organization, and provided technical guidance.

• Optimized molecular cloning techniques for diverse genetic materials production.
• Conducted genetic modification of NK cells via CRISPR/Cas9, mRNA, siRNA, and plasmid-based electroporation (MaxCyte GTx) and develop CAR-NK cells based cell therapy product.
• Managed cancer patient samples, immunophenotyped primary lymphocytes, and conducted functional assays.
• Performing functional assays with modified NK cells against various malignancies using anti-cancer Monoclonal Antibodies (e.g., Daratumumab, Herceptin, Elotuzumab).
• Established gene knockout cell lines using CRISPR/Cas9 technology.

· Developed expertise in histopathological microscopic examination, specializing in Immunohistochemistry (IHC) for cancer research.
· Managed tumor histopathology product lifecycle, optimizing development processes and facilitating collaboration between research entities and R&D teams.
· Established and led teams, spearheaded market setup from inception, and implemented successful sales, marketing, and PR strategies.
· Contributed to product development by creating comprehensive user guides and manuals, ensuring accessibility and usability.
· Enhanced customer satisfaction by resolving technical issues promptly via phone, email, and onsite support, and provided training to facilitate smooth technology adoption.

• Managed mouse models, performing spleen and thyroid extractions, blood collection, and isolation of PBMCs using Ficoll-Paque for FACS staining and immunoassays.
• Isolated T cell subsets via magnetic column-based positive selection methods (LD & MS column, microbeads), conducting studies on immunophenotyping, apoptosis, cell proliferation, activation, and receptor expression.
• Utilized western blotting, PCR, RT-PCR, and ELISA to analyze gene and protein expression kinetics during drug treatment, and conducted non-invasive in-vivo imaging of mice.
• Conducted in-vivo studies on immunomodulation factors using xenografted albino mouse models to investigate the effects of glycolytic inhibitor (2-DG) administration.
• Contributed to ex-vivo studies examining therapeutic responses of cell lines such as J774 under trial conditions.